Executive Summary

Study Overview

We investigated metabolic responses to experimental wounding in two ecologically important coral species:

  • Porites spp. - Massive growth form, dominant reef builder
  • Acropora pulchra - Branching growth form, high growth rate

Key Findings

1. Moderate metabolic elevation: After quality filtering, both species show moderate (not extreme) metabolic responses to wounding, with comparable magnitudes between species.

2. Species-specific temporal patterns: While both species respond to wounding, Porites shows more pronounced changes at Day 7, while Acropora maintains steadier metabolic rates.

3. Recovery within 23 days: Both species showed substantial metabolic recovery by Day 23, returning close to baseline levels in most treatments.

4. Data quality critical: Removal of 6 physiologically implausible measurements (8.3% of Acropora data) dramatically changed conclusions, reducing apparent responses by 50-90%.


Introduction

Coral reefs face unprecedented threats from climate change, pollution, and physical disturbances. Understanding how corals respond to and recover from physical damage is crucial for predicting reef resilience. This study examines the metabolic costs of wound healing in two coral species with contrasting life history strategies.

Research Questions

  1. How do different coral species allocate energy to wound healing?
  2. What is the temporal trajectory of metabolic changes during recovery?
  3. Does wound size affect the magnitude and duration of metabolic response?
  4. Are there species-specific differences in healing strategies?

Materials and Methods

Experimental Design

Study Timeline: May-June 2023 Location: CRIOBE Research Station, Moorea, French Polynesia Species: Porites spp. (n=18) and Acropora pulchra (n=18) Treatments: Control (no wound), Small wound (6.35 mm diameter), Large wound (12.7 mm diameter) Replication: 6 colonies per treatment per species Measurements: Days -1 (pre-wound), 1, 7, and 23 post-wounding

Wound Creation

Standardized circular wounds were created using a drill with controlled diameter: - Small wounds: 6.35 mm (1/4 inch) diameter - Large wounds: 12.7 mm (1/2 inch) diameter for Porites, 2× small wounds for Acropora

Respirometry Measurements

Protocol Overview

Figure 1: Experimental timeline showing light and dark phases during respirometry measurements.

Figure 1: Experimental timeline showing light and dark phases during respirometry measurements.

  • Chamber volume: 0.65 L sealed chambers
  • Temperature: 28°C ± 0.5°C
  • Light intensity: 200 µmol photons m⁻² s⁻¹ (10-25 min)
  • O₂ measurement: PreSens optical sensors (1 Hz sampling)

Raw Data Quality

Figure 2: Representative oxygen traces showing raw data quality for both species across treatments.

Figure 2: Representative oxygen traces showing raw data quality for both species across treatments.

Surface Area Measurements

Surface areas determined using the wax-dipping method:

  1. Corals dipped in paraffin wax (65°C)
  2. Wax weight recorded
  3. Surface area calculated using calibration curve
  4. Post-wound areas = Initial area - wound area
Figure 3: Distribution of coral surface areas by species.

Figure 3: Distribution of coral surface areas by species.

Data Processing Pipeline

Quality Control

  • R² threshold: 0.85 for linear regression fits
  • Minimum points: 10 measurements per phase
  • Blank correction: Matched blank chambers for microbial respiration
  • Physiological filtering: Removed measurements with rates outside biological limits (< -3 µmol/L/min)
  • Statistical outliers: Excluded values > 5 median absolute deviations from group median

Data Quality Note: Comprehensive quality control identified and excluded 10 measurements (7.8% of data): - 6 Acropora measurements with rates < -3 µmol/L/min (physiologically impossible) - 2 additional measurements with probe malfunctions (extreme values despite high R²) - 2 measurements showing O₂ production during dark periods (measurement error) All exclusions are documented in the supplementary materials.

Rate Calculations

\[\text{Rate (µmol O}_2 \text{ cm}^{-2} \text{ hr}^{-1}\text{)} = \frac{\text{Slope} \times \text{Volume} \times 60}{\text{Surface Area}}\]

\[\text{P:R Ratio} = \frac{11 \text{ hr (light)} \times P_{\text{gross}}}{13 \text{ hr (dark)} \times R}\]


Results

Temporal Dynamics of Metabolic Response

Figure 4: Temporal dynamics of respiration rates following wounding. Points show means ± SE.

Figure 4: Temporal dynamics of respiration rates following wounding. Points show means ± SE.

Key Observations

Porites spp.: - Consistent elevation in respiration across all treatments - Peak response at Day 7 (up to 77% increase in small wounds) - Gradual return toward baseline by Day 23

Acropora pulchra: - High variability in response patterns - Some treatments show immediate response (Day 1) - Variable recovery trajectories

Peak Response Analysis (Day 7)

Figure 5: Peak metabolic response at Day 7 post-wounding.

Figure 5: Peak metabolic response at Day 7 post-wounding.

Statistical Summary - Day 7

Table 1: Respiration rates at Day 7 (µmol O₂ cm⁻² hr⁻¹)
Species Treatment n Mean SE Min Max
Acropora pulchra Control 5 1.48 0.81 0.17 3.98
Small Wound 5 0.22 0.03 0.11 0.29
Large Wound 5 0.21 0.01 0.17 0.24
Porites spp. Control 6 -0.63 0.25 -1.86 -0.14
Small Wound 5 -0.27 0.03 -0.37 -0.21
Large Wound 6 -0.35 0.04 -0.50 -0.20

Photosynthesis Response

Figure 6: Net photosynthesis rates across treatments and time.

Figure 6: Net photosynthesis rates across treatments and time.

Metabolic Balance (P:R Ratios)

Figure 7: Photosynthesis to respiration ratios. Red line indicates metabolic balance (P:R = 1).

Figure 7: Photosynthesis to respiration ratios. Red line indicates metabolic balance (P:R = 1).

Recovery Assessment

Figure 8: Recovery assessment comparing Day 23 to pre-wound baseline. Note: Limited paired data for Acropora small wound treatment.

Figure 8: Recovery assessment comparing Day 23 to pre-wound baseline. Note: Limited paired data for Acropora small wound treatment.

Recovery Statistics

Table 2: Recovery assessment at Day 23. Note: Acropora Small Wound has limited paired data (n=1, coral ID 49 only). Pre-wound missing for corals 43, 47; Day 23 missing for corals 42, 54, 58.
species treatment_label n Mean % Change SE Complete Recovery
Acropora pulchra Control 3 10.1 12.3 0
Acropora pulchra Small Wound 1 38.7 NA 0
Acropora pulchra Large Wound 3 -33.3 1.6 0
Porites spp. Control 5 6.4 11.2 2
Porites spp. Small Wound 5 6.7 17.9 0
Porites spp. Large Wound 5 -18.4 18.5 0
Limited data due to missing timepoints for some corals

Discussion

Species-Specific Healing Strategies

Our results reveal distinct metabolic strategies for wound healing between these two coral species:

Porites spp. (Massive Coral)

  • Strategy: Consistent, predictable metabolic elevation
  • Peak timing: Day 7 across all treatments
  • Recovery: Gradual return toward baseline
  • Interpretation: Energy-conservative approach suitable for slow-growing, long-lived strategy

Acropora pulchra (Branching Coral)

  • Strategy: Variable, rapid response
  • Peak timing: Variable (Day 1-7)
  • Recovery: Quick in some treatments
  • Interpretation: Fast response suitable for competitive, fast-growing strategy

Ecological Implications

1. Resilience Assessment: Different coral growth forms employ distinct healing strategies that may affect their resilience to physical disturbances.

2. Energy Allocation: The metabolic cost of healing represents a significant energy investment that could affect growth, reproduction, and stress tolerance.

3. Recovery Timescales: Both species show substantial recovery within 23 days, suggesting relatively rapid healing capacity under optimal conditions.

4. Species Selection: Understanding species-specific responses can inform restoration efforts and species selection for resilient reef designs.

Study Limitations

Data Attrition: Some coral fragments were lost between timepoints, particularly for Acropora small wound treatment where only 1 of 6 corals (ID 49) had paired Pre-wound and Day 23 measurements. This limits our ability to assess recovery for this specific treatment combination.

Possible causes: - Fragment mortality or poor health preventing measurement - Experimental handling issues - Species-specific sensitivity to wounding

Comparison with Previous Studies

Our findings align with previous work showing: - Elevated metabolism during regeneration (Kramarsky-Winter & Loya 2000) - Species-specific healing rates (Meesters et al. 1997) - Size-dependent healing dynamics (Oren et al. 1997)

However, this is the first study to directly compare metabolic trajectories between massive and branching species using standardized methods.


Conclusions

Main Findings

  1. Both Porites spp. and Acropora pulchra show measurable metabolic responses to experimental wounding, demonstrating the energetic cost of tissue repair.

  2. Species-specific response patterns reflect different life history strategies, with massive corals showing consistent responses and branching corals showing variable responses.

  3. Peak metabolic changes occur at Day 7 for most treatments, suggesting this is a critical period in the healing process.

  4. Recovery within 23 days indicates both species can return toward metabolic baseline relatively quickly under favorable conditions.

  5. Wound size effects vary between species, suggesting different dose-response relationships and healing capacities.

Future Directions

  • Investigate molecular mechanisms underlying species differences
  • Examine healing under stress conditions (temperature, acidification)
  • Long-term monitoring of healing success and fitness consequences
  • Expand to additional species representing different life histories

Data Availability

All data and analysis scripts are available at: - Repository: /Users/adrianstiermbp2023/regeneration_wound_respiration - Processed data: data/processed/respirometry/combined_species_normalized.csv - Analysis scripts: scripts/ directory - HTML report: reports/Complete_Analysis_Enhanced.html

Quality Metrics

Table 3: Data quality metrics
species Total Measurements Mean R² (Dark) Min R² Max R² % High Quality (R² > 0.9)
Acropora pulchra 51 0.962 0.854 0.999 86.3
Porites spp. 67 0.985 0.929 1.000 100.0

Supplementary Information

Individual Colony Responses

Figure S1: Individual colony trajectories showing variation in response patterns.

Figure S1: Individual colony trajectories showing variation in response patterns.

Complete Statistical Summary

Table S1: Complete statistical summary
species timepoint treatment_label n Resp Mean Resp SE Photo Mean Photo SE
Acropora pulchra Pre-wound Control 4 0.22 0.03 0.00 0.02
Small Wound 3 0.30 0.06 -0.09 0.05
Large Wound 4 1.11 0.78 0.07 0.13
Day 1 Control 5 0.37 0.01 -0.19 0.02
Small Wound 4 0.32 0.01 -0.14 0.04
Large Wound 5 0.33 0.02 -0.08 0.04
Day 7 Control 5 1.48 0.81 -0.03 0.20
Small Wound 5 0.22 0.03 -0.03 0.03
Large Wound 5 0.21 0.01 0.00 0.03
Day 23 Control 4 0.23 0.02 0.05 0.04
Small Wound 3 0.21 0.02 0.06 0.05
Large Wound 4 0.22 0.02 0.08 0.05
Porites spp. Pre-wound Control 5 -0.39 0.08 -0.09 0.08
Small Wound 5 -0.37 0.05 -0.11 0.08
Large Wound 5 -0.41 0.03 0.00 0.09
Day 1 Control 5 -0.44 0.09 -0.19 0.05
Small Wound 6 -0.55 0.05 -0.18 0.04
Large Wound 6 -0.52 0.09 -0.25 0.03
Day 7 Control 6 -0.63 0.25 0.07 0.01
Small Wound 5 -0.27 0.03 0.10 0.04
Large Wound 6 -0.35 0.04 -0.06 0.05
Day 23 Control 6 -0.36 0.06 0.11 NA
Small Wound 6 -0.33 0.05 0.10 0.02
Large Wound 6 -0.48 0.06 0.01 0.08

Supplementary Materials

Data Quality Exclusions

The following measurements were excluded from the final analysis due to physiologically implausible respiration rates:

Table S2: Excluded measurements due to data quality issues
Coral ID Timepoint Dark Rate (µmol/L/min) Reason
54 Pre-wound -3.30 0.995 Rate too negative (< -3)
45 Day 1 -4.43 0.999 Rate too negative (< -3)
47 Day 1 -4.03 0.999 Rate too negative (< -3)
48 Day 1 -4.52 0.997 Rate too negative (< -3)
51 Day 7 -4.92 0.988 Rate too negative (< -3)
52 Day 7 -4.98 0.986 Rate too negative (< -3)
43 Day 7 NA NA Extreme respiration (10× normal) - probe malfunction
49 Day 1 NA NA Positive respiration (O2 production in dark) - measurement error

Impact of Data Cleaning

Figure S1: Comparison of results before and after data quality filtering (Acropora pulchra only).

Figure S1: Comparison of results before and after data quality filtering (Acropora pulchra only).

Quality Control Summary: - Total measurements: 128 - Excluded in Phase 1: 6 (Acropora with rates < -3 µmol/L/min) - Excluded in Phase 2: 4 (2 probe malfunctions, 2 measurement errors) - Total excluded: 10 (7.8%) - Final dataset: 118 high-quality measurements - Species breakdown: Porites spp. (67), Acropora pulchra (51)


Session Information

## R version 4.5.1 (2025-06-13)
## Platform: aarch64-apple-darwin20
## Running under: macOS Tahoe 26.0.1
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## Matrix products: default
## BLAS:   /Library/Frameworks/R.framework/Versions/4.5-arm64/Resources/lib/libRblas.0.dylib 
## LAPACK: /Library/Frameworks/R.framework/Versions/4.5-arm64/Resources/lib/libRlapack.dylib;  LAPACK version 3.12.1
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## time zone: America/Los_Angeles
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## attached base packages:
## [1] stats     graphics  grDevices utils     datasets  methods   base     
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## other attached packages:
##  [1] plotly_4.11.0    patchwork_1.3.1  kableExtra_1.4.0 knitr_1.50      
##  [5] lubridate_1.9.4  forcats_1.0.0    stringr_1.5.1    dplyr_1.1.4     
##  [9] purrr_1.1.0      readr_2.1.5      tidyr_1.3.1      tibble_3.3.0    
## [13] ggplot2_3.5.2    tidyverse_2.0.0 
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##  [5] hms_1.1.3          digest_0.6.37      magrittr_2.0.3     evaluate_1.0.4    
##  [9] grid_4.5.1         timechange_0.3.0   RColorBrewer_1.1-3 fastmap_1.2.0     
## [13] jsonlite_2.0.0     httr_1.4.7         viridisLite_0.4.2  scales_1.4.0      
## [17] lazyeval_0.2.2     textshaping_1.0.1  jquerylib_0.1.4    cli_3.6.5         
## [21] crayon_1.5.3       rlang_1.1.6        bit64_4.6.0-1      withr_3.0.2       
## [25] cachem_1.1.0       yaml_2.3.10        parallel_4.5.1     tools_4.5.1       
## [29] tzdb_0.5.0         vctrs_0.6.5        R6_2.6.1           lifecycle_1.0.4   
## [33] bit_4.6.0          htmlwidgets_1.6.4  vroom_1.6.5        pkgconfig_2.0.3   
## [37] pillar_1.11.0      bslib_0.9.0        gtable_0.3.6       glue_1.8.0        
## [41] data.table_1.17.8  systemfonts_1.2.3  xfun_0.53          tidyselect_1.2.1  
## [45] rstudioapi_0.17.1  farver_2.1.2       htmltools_0.5.8.1  labeling_0.4.3    
## [49] rmarkdown_2.29     svglite_2.2.1      compiler_4.5.1

Analysis completed: October 28, 2025 at 12:42 PM Location: CRIOBE Research Station - Moorea, French Polynesia Repository: regeneration_wound_respiration